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A protein expression system for tandem affinity purification in Xanthomonas citri subsp citri

Texto completo
Dantas, Giordanni C. [1] ; Martins, Paula M. M. [1] ; Martins, Daniela A. B. [2] ; Gomes, Eleni [3] ; Ferreira, Henrique [1]
Número total de Autores: 5
Afiliação do(s) autor(es):
[1] Univ Estadual Paulista, Inst Biociencias, Dept Bioquim & Microbiol, Rio Claro, SP - Brazil
[2] Univ Estadual Paulista, Inst Quim, Dept Bioquim & Tecnol Quim, Araraquara, SP - Brazil
[3] Univ Estadual Paulista, Dept Biol, Sao Jose Do Rio Preto, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: Brazilian Journal of Microbiology; v. 47, n. 2, p. 518-526, APR-JUN 2016.
Citações Web of Science: 0

Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp. citri (Xac), is one of the most devastating diseases to affect citrus crops. There is no treatment for citrus canker; effective control against the spread of Xac is usually achieved by the elimination of affected plants along with that of asymptomatic neighbors. An in depth understanding of the pathogen is the keystone for understanding of the disease; to this effect we are committed to the development of strategies to ease the study of Xac. Genome sequencing and annotation of Xac revealed that similar to 37% of the genome is composed of hypothetical ORFs. To start a systematic characterization of novel factors encoded by Xac, we constructed integrative-vectors for protein expression specific to this bacterium. The vectors allow for the production of TAP-tagged proteins in Xac under the regulation of the xylose promoter. In this study, we show that a TAP-expression vector, integrated into the amy locus of Xac, does not compromise its virulence. Furthermore, our results also demonstrate that the polypeptide TAP can be overproduced in Xac and purified from the soluble phase of cell extracts. Our results substantiate the use of our vectors for protein expression in Xac thus contributing a novel tool for the characterization of proteins and protein complexes generated by this bacterium in vivo. (C) 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. (AU)

Processo FAPESP: 06/59494-9 - Localizacao sub-celular de proteinas marcadas com gfp em xanthomonas axonopodis pv. citri por microscopia de fluorescencia.
Beneficiário:Paula Maria Moreira Martins
Modalidade de apoio: Bolsas no Brasil - Mestrado
Processo FAPESP: 13/50367-8 - New environmental-friendly compounds to combat Citrus canker
Beneficiário:Henrique Ferreira
Modalidade de apoio: Auxílio à Pesquisa - Regular
Processo FAPESP: 04/09173-6 - Caracterização funcional de proteínas hipotéticas de bactérias fitopatogênicas
Beneficiário:Henrique Ferreira
Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
Processo FAPESP: 13/19806-5 - Seleção de sítio de divisão em Xanthomonas citri subsp. citri: caracterização de minCD
Beneficiário:Giordanni Cabral Dantas
Modalidade de apoio: Bolsas no Brasil - Doutorado